Searching biomarker candidates in serum using multidimensional native chromatography. I. Enhanced separation method.
The microplate-based method developed by our group for non-denaturing multidimensional proteome separation was improved on using improved column arrays and a newly developed robot. Currently size exclusion, anion exchange and lectin affinity chromatography are combined orthogonally. Different samples run simultaneously to enhance reliability of intercomparison. LC-ESI (electro-spray ionization) MS/MS analysis of selected fractions identified 32,288 peptides matching 2,669 serum proteins. The present contribution (I) shows the characteristics of the method, whereas "prove of principle" by applying it to search for biomarker candidates with model diseases is reported in an accompanying paper (II).
Kreusch
Stefan
S
Institute of Biochemistry, Friedrich Schiller University, Medical Faculty, D-07740 Jena, Germany.
Schulze
Margarete
M
Cumme
Gerhard A
GA
Ditze
Günter
G
Moore
Thomas
T
Rhode
Heidrun
H
D015415:Biomarkers / Q000097:blood*; D001798:Blood Proteins / Q000032:analysis*; D002845:Chromatography / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D030562:Databases, Protein; D006801:Humans; D010455:Peptides / Q000032:analysis*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods; D015203:Reproducibility of Results; D012371:Robotics / Q000295:instrumentation; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
D015415:Biomarkers; D001798:Blood Proteins; D010455:Peptides